STUDYGUIDE

Species specific module:

Ruminants (goat, sheep, cattle)

Introduction to Laboratory Animal Science

Organisation:
Department Farm Animal Heath
Faculty of Veterinary Medicine Utrecht University

Contact and responsible for Module Ruminants:

G.A. Hooijer (g.a.hooijer@uu.nl 030-2531248)

www.uu.nl/lascourse
www.uu.nl/lasspecies

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Contents
Objective of the module...................................................................................................... 3
Competence ......................................................................................................................... 3
Workload ............................................................................................................................... 3
Course material ................................................................................................................... 3
Learning outcomes ............................................................................................................. 4
Final examination ................................................................................................................ 4
Certificate.............................................................................................................................. 4
Program outline* .................................................................................................................. 5
1. Introduction ruminant course .......................................................................................... 5
2. Animal Handling ............................................................................................................. 6
3. Sedation, analgesia, anaesthesia ..................................................................................... 7
4. Importance of diseases of ruminants in laboratory conditions ....................................... 8
5. General Anatomy, with special focus on Cattle, Sheep and Goats ................................. 9
6. Physiology....................................................................................................................... 9
7. Housing and care of ruminants in laboratory conditions .............................................. 10
8. Practical Basic Techniques ........................................................................................... 11
9. Demonstration Basic Techniques ................................................................................. 11
10. LAS assignment ............................................................................................................ 12
General Anatomy, with special focus on Cattle, Sheep and Goats .......................... 13

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Objective of the module
The objective of this module is to present basic and appropriate biology, care, health
and management of ruminants, recognition of pain, suffering and distress in these
animals and minimally invasive procedures without anaesthesia to be applied on
these animals. This course meets the standards for the species-specific education
and training requirements for persons designing projects and procedures for
ruminants.

Competence
For the function ‘designing procedures and projects’ the Dutch government requires
competency.
On December 18, 2014 the new legislation regarding animal experimentation was
implemented. Due to this implementation the course on Laboratory Science has
been adapted. The course contains now two parts, a basic course (Introduction to
laboratory animal science) and a species specific module. The basic course
certificates and, at least, one species specific certificate will give the required
competence.
From August 1, 2015 the competence is limited. You are not allowed to perform any
procedure on animals, unless there is supplementary education. From now you are
competent when you are skilled (competence profile, species and skill(s) stated).
After successful completion of the new basic course and after successful completion
of the present module on ruminants, you will be competent to design procedures and
projects and to execute simple procedures on these animals. Further skills have to
be obtained by working under supervision until competence is demonstrated. Only
then are you allowed to work with animals independently.

Workload
The number of credits that can be obtained after successful completion of the course
is 1.4 ECTS (European Credit Transfer and Accumulation System). The length of the
course is one week (fulltime, 09.00-17.30 hrs). Participation in all parts of the course
is mandatory. Knowledge of the content of course material is necessary at the start
of the course.

Course material
During the course the book Principles of Laboratory Animal Science, revised edition,
2001, L.F.M. van Zutphen, V. Baumans and F. Ohl (eds.) (ISBN 13: 978-0-444-
50612-2) will be used. Bibliography given below. Further material will be provided on
a memory-stick.

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Learning outcomes
Learning outcomes are in accordance with the EC Training and Education Document
relating to Directive EU/2010/63,
http://ec.europa.eu/environment/chemicals/lab_animals/pdf/Endorsed_E-T.pdf
The species-specific course includes the following modules:
3.1 Basic and appropriate biology
3.2 Basic and appropriate biology – species specific (practical)
4. Animal care, health and management (theory)
5. Recognition of pain, suffering and distress – species specific
6. Humane methods of killing (theory)
7. Minimally invasive procedures without anaesthesia (theory)
8. Minimally invasive procedures without anaesthesia – species specific (skills).
For details, see separate chapters in the study guide.

Final examination
As a group you have to analyze a given article with help of a questionnaire that is
based on the ARRIVE guidelines: (Kilkenny C, Browne WJ, Cuthill IC, Emerson M,
Altman DG (2010) Improving bioscience research reporting: the ARRIVE guidelines
for reporting animal research.). Based on every aspect you learned about during the
course you have to describe in detail how the experiments described in the article
are performed. You will report this in a small presentation (10-15 min) at the end of
the course and discuss your choices with the critical audience- the other students.
Each student gets a mark ranging from 0-10 points (0= unsatisfactory, 10=very
good) depending on the quality. The examination involves the handbook, the
lectures, discussions, demonstrations and practical’s.

Certificate
If the participant has met all legal requirements and has successfully passed the
exam at the end of the course, the participant will receive a certificate for this species
specific module Ruminants.
If you do not meet all legal requirements, like not holding a Master degree (yet),
you will receive a written confirmation stating that you followed the module and
successfully passed the exam. Together with the certificate for the basic course,
after obtaining your Master degree, you can request the final certificate confirming
that you meet all legal requirements of the Dutch legislation to be registered for the
function “designing projects and procedures for ruminants”.

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Program outline*

Title 1. Introduction ruminant course

Lecturers Lecturers Ruminant Health Institute
Content During this interactive lecture an overview of all topics to be
presented during the entire course period will be given. The current
national legislation governing the use of ruminants in research will be
introduced. In dependence on the natural social structure of these
animals implications for housing and testing of these animals will be
discussed. Special attention will be paid to welfare related questions.
Key Text a. Cattle
b. http://www.cowsignals.com/books/cow_signals_-
_english_edition
c. http://www.cowsignals.com/books/from_calf_to_heifer_-
_english_edition
d. Information selected from the following books:
http://www.cowsignals.com/books/dry_period_special_needs_
cows_-_english_edition
http://www.cowsignals.com/books/udder_health_-
_english_american_edition
http://www.cowsignals.com/books/feeding_signals_-
_english_edition
http://www.cowsignals.com/books/cow_signals_checkbook_-
_english_edition
e. Sheep
f. http://www.roodbont.nl/en/sheep_signals_concept/index.php
g. Goat
h. https://www.rspca.org.uk/ImageLocator/LocateAsset?asset=d
ocument&assetId=1232713000349&mode=prd

Form Lecture
Testing your 1. Describe the life cycle of a common dairy cow, sheep, goat
competence 2. Describe the most essential needs for cattle, sheep, goats used
in experiments
3. When would you decide on cow / calve , sheep or goat as model
organism?
4. Give an example of how ruminants might be used as model to
study human disease.

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Title 2. Animal Handling
Lecturer Lecturers Ruminant Health Institute
Content In this setting you will be made able to restrain ruminants. This
practicum will prepare you for skills on animal handling. Important to
understand that 95% of a procedure is restraining the animal in the
right way and a quiet behavior. Restraining is handling an animal in
such a way that nor you neither the animal will be harmed when
performing a technique.
Working safely and concern for animal welfare with laboratory
animals requires a core competency in handling and restraint
methods. Proper restraint and handling techniques are essential for
clinical examination and injections while limiting the amount of
stress and/or discomfort to the animal and to the handler.
Training will be done in the cubicle stall. You will be taught how to
approach, tether a halter, escort, restrain, check ( take temperature,
check gait, check claws of cattle and small ruminants etc.) of the
ruminants. In the first place concerning welfare of the animals.

Key Text a) http://www.nfacc.ca/codes-of-practice/beef-cattle

b) http://www.nfacc.ca/codes-of-practice/dairy-cattle
c) http://www.nfacc.ca/pdfs/codes/goat_code_of_practice.pdf
d) http://www.nfacc.ca/pdfs/codes/sheep_code_of_practice.pdf
Form Lecture and practicum
Testing your Describe every own specific way of handling and restraining cattle,
competence sheep and goats.
Describe possible consequences (for the animal and for the
handler) if animals are not handled and/or restraint properly.
Demonstrate certain skills in animal handling

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Title 3. Sedation, analgesia, anaesthesia
Lecturers Dep. Gezondheidszorg Paard – Afd. Veterinaire Anesthesiologie
/ Lecturers Ruminant Health Institute
Content This lecture attempts to provide a basic level of knowledge in
sedation, analgesia and anesthesia for research workers with
special emphasis ruminants in laboratory conditions. The
pathophysiology of pain and knowledge concerning different
techniques providing analgesia are reviewed. Drugs for either
local, regional and total anaesthesia will be discussed. The
general principles of peri-operative care, anesthetic techniques
and anesthetic management are discussed including injection-
and inhalation anesthesia.
The most commonly used ways of anaesthesia are interactively
demonstrated:
 intaveneous (jugular-, mammary vein, ear vein = vena
auricularis lateralis / vena auricularis intermedia, tail vein
= v. sacralis mediana, foot vein = v. digitalis palmaris /
communis)
 local, regional (paravertebral, corneal nerve, lacrimal
nerve, inverted l-block, epidural, spinal)
 films of these methods used in practice
Key Text Cattle:
https://www.acvs.org/files/proceedings/2012/data/papers/169.pdf
Small ruminants:
https://www.acvs.org/files/proceedings/2012/data/papers/170.pdf

Form (Interactive) lecture

Testing 1. Describe pre-emptive analgesia
your 2. Describe the pillars of anesthesia
competence 3. Describe use of regional anaesthesia in ruminants

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Title 4. Importance of diseases of ruminants in
laboratory conditions
Lecturer Lecturers Ruminant Health Institute
Content Diseases in animals can be a consequence of experimental
setup or occur as an unintended effect during the study. This
lecture will focus on how to handle when animals have aberrant
status praesens. In particular, the risks of disease at different
levels will be discussed as well as how to recognize disease and
come to a diagnosis. Furthermore, the most common
(infectious) diseases of ruminants are reviewed, with a special
focus on zoonosis. ( Brucellosis, Listeriosis, Leptospirosis,
Salmonellosis, Microsporum, trichophyton, Sarcoptes,
Campylobacter, Chlamydophilia, Ecthyma contagiosum, Johne’s
disease )
Measurements to reduce the infection risks as well as how to
handle in case of infection the potential effects on the studies.
Key Text  Confronting Zoonoses, Linking Human and Veterinary
Medicine; Kahn, Emerging Infectious Diseases, 2006
 Code of Practice: Welzijnsbewaking van proefdieren
 Robbins & Cotran Pathologic Basis of Disease, 9e (for
reference only)
 http://www.vet.cornell.edu/consultant/consult.asp
Form Interactive lecture
Testing your 1. How to handle when laboratory animals are ill or diseased?
competence 2. What are actions with regard to pathology, clinical and
microbiological diagnostics that can be considered?
3. What can be consequences of diseases in laboratory
animals?
4. What is the importance of collection of samples and how can
they be used to diagnose disease?
5. Give an example of how a natural pathogen in lab animals
can interfere with research.
6. List the transmission routes of pathogens and indicate what
measurements can be taken to reduce the risks of
spreading.

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Title 5. General Anatomy, with special focus on
Cattle, Sheep and Goats
Lecturer Lecturers Ruminant Health Institute
Content ‘One of the prerequisites for responsible use of animals in
biomedical research is a thorough knowledge of the biological
characteristics of the species to be used. The choice of animal
species, sex, age and strain largely depend on whether their
anatomical characteristics are suited to the research demands’.

In this lecture the general anatomy of mammals will be

discussed. Special attention will be given to the anatomy of
ruminants in relation to biomedical research. A visit to the
Anatomy Department will clarify interspecies differences
Key Text Principles of laboratory animal science, chapter 3
‘Biology and husbandry of laboratory animals’.
Handboek Proefdierkunde, hoofdstuk 3; Biologie en Zoötechniek
Form Lecture
Testing your Describe important differences in the gastrointestinal tract in
competence young and older ruminants

Title 6. Physiology
Lecturer Lecturers Ruminant Health Institute
Content Stress, defined as any deviation from homeostasis, induces a
behavioral and physiological response that may impact on a
wide array of animal experiments. In this lecture, the basic
principles of homeostasis and physiological response to stress
will be discussed. Specific topics that will be addressed are:
- Autonomic nervous system and sympathetic and
parasympathetic response
- Impact on cardiovascular system
- Neuroendocrine response to stress
- Impact on animal studies
Key Text Handbook “Principles of Laboratory Animal Science” (course
book), chapter “Behavior, stress and welfare”, Environment and
Physiology.
Physiology and anatomy in relation to make a decision tree
for finding the optimal animal model.
Form Lecture
Testing your How is stress defined?
competence Which physiological systems are affected in response to stress?
What is the impact of a sympathetic and neuroendocrine
response to stress?
Why is it important to consider stress in animal experiments?

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 7. Housing and care of ruminants in
laboratory conditions
Lecturer Lecturers Ruminant Health Institute
Content In this interactive working group assignment students and
lecturer will discuss housing and care of ruminants in laboratory
conditions, assessment of discomfort, clinical signs indicating
the humane endpoint and the best method to euthanize animals
(least distress and not interfering with experimental results). On
the basis of three hypothetical experiments the above
mentioned topics will be discussed: 1. Accommodation of the
animals 2. How to assess discomfort 3. Selection of the most
appropriate method of euthanasia
Films about these issues will be demonstrated
Key Text  “Principles of Laboratory Animal Science; chapter 4
(Behavior, stress and wellbeing)
 Council of Europe Convention ETS 123 for the protection of
vertebrate animals used for experimental and other scientific
purposes. Appendix A: Guidelines for accommodation and
care of animals.
 EU Working Party Report: Recommendations for euthanasia
of experimental animals Parts 1&2
 EU Directive 2010/63/EU on the protection of animals used
for scientific purposes; Annex IV: Methods of killing animals
 Key text mentioned in ‘Introduction’
Form Interactive working group assignment
Testing your For each of the three hypothetical experiments
competence 1. What is the most appropriate way to house ruminants in
laboratory conditions?
2. How do you assess distress, pain in ruminants in laboratory
conditions?
3. What is the most appropriate method to euthanize ruminants
in laboratory conditions?
4. Which clinical symptoms should be used to define the
(humane) endpoint of the animal experiments?
5. In general: how do you prevent avoidable discomfort in
ruminants in laboratory conditions?

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Title 8. Practical Basic Techniques
Lecturer Lecturers Ruminant Health Institute
Content This practical contains a brief description of some basic
procedures concerning the administration of drugs. Dutch law
demands responsible drug use in animal husbandries in
general and especially usage of antibiotics. Possible
associations between antimicrobial use and the prevalence of
resistance of micro-organisms are discussed. Basic principles
of surgery and some administration procedures will be shown
and practised. Administering substances to animals, for
whatever reason, can have a significant impact on their
welfare. If carried out incorrectly, not only can animal welfare
be compromised, but the scientific goals of study can be
affected. In practice participants will be instructed in the
administration of substance (injections i.m., s.c., i.v., oral,
pour-on) to ruminants.
Films about these issues will be demonstrated
Key Text a) Handbook “Principles of Laboratory Animal Science”
(course book), chapter 16 “Experimental procedures”.
b) Videos on usb-stick on handling/restraining
c) http://www.autoriteitdiergeneesmiddelen.nl/en/home
d) https://www.youtube.com/watch?v=JCwwBEQrMRI
e) https://www.youtube.com/watch?v=i3wrvib1ENY
f) https://www.youtube.com/watch?v=m0TFBU75DiM
g) https://www.youtube.com/watch?v=m5DtF3QO710
h) https://www.youtube.com/watch?v=iSZEyRoeT4k
Form Lecture and practical
Testing your Which methods of administration do you know?
competence Which routes of administration do you know?
How do you decide on a certain method of administration?
Understanding a surgical procedure and the different steps
needed.

Title 9. Demonstration Basic Techniques

Lecturer Lecturers Ruminant Health Institute
Content In this demonstration a brief description of some basic
procedures concerning sampling (blood, abdominal fluid etc),
feces and urine will be discussed. Injections s.c., i.m., i.v. and
the practical ways to take blood samples will be
demonstrated/done
Key Text Videos on memory-stick on handling/restraining
Form Lecture and demonstration
Testing your Which methods of blood sampling do you know?
competence How do you decide on a certain sampling technique ?
When would you anesthetize your animals for sampling and
when not? How would you sample urine and feces of mice?
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 How would you do that when animals are kept in groups?

Title 10. LAS assignment

Lecturers Dr. G.A. Hooijer : g.a.hooijer@uu.nl; Dr. R. Jorritsma
(2 per course) r.jorritsma@uu.nl
Content As a group you have to analyze a given article with help of a
questionnaire that is based on the ARRIVE guidelines: (Kilkenny
C, Browne WJ, Cuthill IC, Emerson M, Altman DG (2010)
Improving bioscience research reporting: the ARRIVE guidelines
for reporting animal research.). Based on every aspect you learned
about during the course you have to describe in detail how the
experiments described in the article are performed. You will report
this in a small presentation (10-15 min) at the end of the course
and discuss your choices with the critical audience- the other
students.
Key Text  Van Zutphen, L.F.M., Baumans, V., Beynen A.C. (2001)
"Principles of Laboratory Animal Science: A Contribution to the
Humane Use and Care of Animals and to the Quality of Experi-
mental Results"
 Additional literature (sought by the participants themselves)
Form Working group assignment

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Schedule species specific course Ruminants

Time Subject Room Teacher

Day 1 9.00-10.15 Introduction ruminant course Gerrit Hooijer
10.30-12.30 General Anatomy, with special focus on Paul Heijnen
Cattle, Sheep and Goats

13.15-14.45 Physiology Marieke Lindhout-

Dijkshoorn
15.00-16.30 Animal Handling Dick Scholten

Day 2 9.00-10.30 Self-study as preparation on diseases in

ruminants
10.45-12.30 Importance and / of diseases of ruminants in Gerrit Hooijer
laboratory conditions

13.00-14.00 Preparation: housing systems in ruminants.

Welfare and needs
14.15-16.30 Housing and care of ruminants in laboratory Hilde Aardema
conditions

Day 3 9.00-12.30 Sedation, analgesia, anaesthesia Janny de Grauw

13.00-15.00 Practical Basic Techniques Dick Scholten

15.00-16.00 Demonstration Basic Techniques Dick Scholten
Day 4 9.00-12.30 Preparation presentation or room for
additional individual program

13.15-17.00 Presentation articles Gerrit Hooijer

Day 5 9.00-11.30 LAS assignment Gerrit Hooijer /

Ruurd Jorritsma

11.45-12.30 Course evaluation Gerrit Hooijer

* The schedule as mentioned above is intended to provide an overview of the course contents. Exact times 
and locations  for the individual lectures, practicals and working groups may vary from course to course, 
dependent on the availability of the teachers and will be made available prior to the start of each course 

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Modules art. 23.2.b function Species specific
Module 3.1: Basic and appropriate biology – species specific (theory)

This module provides an introduction to the basic principles of animal behaviour,

care, biology and husbandry. It incorporates information in relation to anatomy and
physiological features, including reproduction, behaviour and routine animal
husbandry and enrichment practices. It is not intended to provide more than the
minimum background information which is needed for someone to be able to begin
work under supervision.
Following this module practical training, under supervision, should provide each
individual with the expertise and skills needed for them to carry out their particular
function. Practical training requirements will, inevitably, differ according to function.

Learning Outcomes
Trainees should be able to:
3.1.1. Describe basic anatomy, physiology, reproduction and behaviour of the
relevant species.
3.1.5. Describe the dietary requirements of the relevant animal species and explain
how these can be met.
3.1.6. Describe the importance of providing an enriched environment (appropriate to
both the species and the science) including social housing and opportunities for
exercise, resting and sleeping.
3.1.7. When relevant to the species, recognise that there are different strains, and
that these can have different characteristics which can affect both welfare and
science.
3.1.8. When relevant to the species, recognise that alterations to the genome can
affect the phenotype in unexpected and subtle ways, and the importance of
monitoring such animals very carefully.
3.1.9. Maintain and interpret accurate, comprehensive records of animals held in the
animal facility, including the wellbeing of the animals

Module 4: Animal care, health and management – species specific (theory)

This module provides information on various aspects of animal health, care and
management including, environmental controls, husbandry practices, diet, health
status and disease. It also includes relevant basic learning outcomes relating to
personal health and zoonoses.

Learning Outcomes
Trainees should be able to:
4.1. Describe suitable routines and husbandry practices for the maintenance, care
and welfare for a range of animals used in research, to include small laboratory
species and large animal species where appropriate.
4.2. Describe suitable housing conditions for laboratory animals, how conditions are
monitored and identify the consequences for the animal resulting from inappropriate
environmental conditions.
4.6. Describe how to provide water and an appropriate diet for laboratory animals
including the sourcing, storage and presentation of suitable foodstuffs and water

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4.7. List the methods, and demonstrate an understanding of appropriate, safe and
humane handling, sexing and restraint of one or more named species for common
scientific procedures.
4.8. Name different methods for marking individual animals and state an advantages
and disadvantage for each method.

4.9. List potential disease risks in the animal facility, including specific predisposing
factors which may be relevant. Name methods available for maintaining appropriate
health status (including use of barriers, different containment levels use of sentinels
as relevant to the species).
4.10. Describe appropriate breeding programmes
4.11. Describe how genetically altered animals can be used for scientific research
and the importance of monitoring such animals very carefully.
4.12. List the correct procedures for ensuring health, welfare and care of animals
during their transport.

Module 5: Recognition of pain, suffering and distress - species specific

This module prepares individuals to be able to identify normal condition and

behaviour of experimental animals and enable them to differentiate between a
normal animal and one which is showing signs of pain, suffering or distress which
could be a result of factors including environment, husbandry or the effect of
experimental protocols. It will also provide information regarding severity
classifications, cumulative severity and the use of humane endpoints.

Learning Outcomes
Trainees should be able to:
5.1. Recognise normal or desirable behaviour and appearance of the individuals in
the context of species, environment and physiological status.
5.2. Recognise and explain the origin of abnormal behaviour and signs of discomfort,
pain, suffering, or distress, as well as signs of positive well-being and principles of
how pain, suffering and distress can be managed.
5.3. Discuss factors to be considered and methods available for assessing and
recording the welfare of animals e.g. score sheets.
5.4. Describe what a humane end point is. Identify criteria to be used to set humane
endpoints. Define action to be taken when a humane endpoint is reached and
consider possible options for refining methods to finish at an earlier endpoint.
5.5. Describe the severity classifications included in the Directive and give examples
of each category; explain cumulative severity and the effect this may have on the
severity classification.
5.6. Describe the circumstances when anaesthesia or analgesia may be necessary
to minimise pain, suffering, distress or lasting harm

Module 7: Minimally invasive procedures without anaesthesia – species

specific (theory) [Function Specific for Functions A and B]

This module provides an introduction to the theory relating to minor procedures. It

provides information about appropriate methods of handling and restraint and
describes appropriate techniques for injection, dosing and sampling relevant to the
species. It should provide information sufficient for individuals to understand what will

15
be required of them before they go on to trained in the practical aspects of these
skills whilst under supervision.

Learning Outcomes
Trainees should be able to:
7.1. Describe appropriate methods and principles to be followed when handling
animals (including methods of manual restraint and use of restricted environments).
7.2. Describe the biological impact of procedures and restraint on physiology.
7.3. Describe refinement opportunities for procedures and restraint
7.4. Describe techniques/procedures including, for example, injection, sampling and
dosing techniques (routes/volumes/frequency), dietary modification, gavage, tissue
biopsy, behavioural tests, use of metabolic cages.
7.5. Describe how to perform minor techniques (without anaesthesia) and relate
appropriate sample volumes and sampling frequencies for the relevant species.
7.6. Describe the need for rigour and consistency in conducting scientific procedures
and the correct recording and handling of samples.
7.7. Describe appropriate methods for the assessment of the welfare of animals with
respect to the severity of procedures and know what appropriate action to take.
7.8. Recognize that refinement is an on-going process and know where to find
relevant, up-to-date, information.
7.9. Describe the biological consequences of transport, acclimatization, husbandry
conditions and experimental procedures on the species concerned and describe how
these can be minimised.

Module 8: Minimally invasive procedures without anaesthesia – species

specific (skills) [Function Specific for Function A]

This module delivers practical elements of training relevant to Module 7. Practical

training for minor procedures can be taught through a number of methods using
different tools which are available and designed for the purpose (this is likely to
include synthetic animal models and the use of cadavers). The module should be
designed in such a way that it will enable the trainee to attain a level of proficiency
such that, when commencing work under supervision, s/he should cause no pain,
suffering, distress or lasting harm to the animal.

Learning Outcomes
Trainees should be able to:
8.1. Select and explain the best methods for common procedures (such as blood
sampling and application of substances) including route/volume/ frequency as
appropriate.
8.2. Demonstrate that s/he can handle and restrain the animal in the best position for
the technique.
8.3. Perform minor techniques under supervision, in a manner that does not inflict
unnecessary pain, suffering, distress or lasting harm.

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